Von Willebrand Activity Research Articles

Factor VIII related antigen in platelets of patients with Von Willebrand's disease

Presence of Factor VIII related antigen (VIII R:Ag) associated with Von Willebrand activity (VIII R:WF) in platelets has been reported. According to the literature, the situation in patients with Von Willebrand's disease (VWd) is not clear. A new collaborative study was undertaken concerning 17 patients with well documented VWd. The study included several homozygotes or double heterozygotes and genetic variants of the disease. The results showed that severely affected patients with undetectable VIIIR:Ag in their plasma had no detectable VIII R:Ag in their platelets, all other patients had detectable amounts of antigen in their platelets some of them in higher concentration than normals.

Inhibition of human platelet aggregation by the proteolytic effect of streptokinase. Role of the human factor-VIII-related protein.

Defective ADP-induced aggregation was observed in in vitro streptokinases(SK)-treated normal platelet-rich plasma. Classic haemophilia and normal platelet poor plasma (PPP) treated with SK inhibit the aggregation of washed platelets; plasmin-treated normal human serum also shows an inhibitory effect on platelet aggregation. However, SK-treated von Willebrand plasmas do not inhibit the aggregation of washed platelets. This confirms the fact that the anti-aggregating effect is mainly linked to the digested factor VIII) but not to the digested fibrinogen. Defective ristocetin-induced platelet aggregation has also been observed in SK-treated plasmas. The presence of normal PPP does not modify the inhibition of the ADP-induced aggregation of washed platelets in SK-treated PPP. However, it does correct the ristocetin-induced aggregation. These results suggest that the inhibition of the ADP-induced aggregation is caused by the factor VIII degradation products, while the inhibition of the ristocetin-induced aggregation appears because of a defective von Willebrand activity of the factor VIII molecule.

Molecular structural studies of human factor VIII.

Neither normal nor hemophilic factor VIII protein enters a 5% sosium dodecyl sulfate gel; on reduction, however, a single 195 000-molecular-weight peptide is observed. Hemophilic and normal factor VIII contain carbohydrate and appear identical in subunit molecular weight, electrical charge, and major antigenic determinants. Thrombin activation and inactivation of factor VIII does not detectably change the subunit molecular weight. Trypsin causes similar activity changes and obviously cleaves the factor VIII subunit. Human plasmin destroys factor VIII procoagulant activity and degrades the factor VIII subunit to 103 000-, 88 000-, and 17 000-molecular-weight peptides. Both normal and hemophilic factor VIII as well as thrombin-inactivated factor VIII support ristocetin-induced platelet aggregation. Purified factor VIII chromatographed on 4% agarose in 1.0 M sodium chloride shows no dissociation of the procoagulant activity from the void volume protein. Gel chromatography on 4% agarose in 0.25 M calcium chloride results in a procoagulant activity peak removed from the void volume protein; both peaks contain protein which does not enter a 5% SDS gel, but on reduction a 195 000-molecular-weight subunit band is observed for each. Both the void volume protein peak and the procoagulant activity peak from the 0.25 M calcium chloride-agarose gel column support ristocetin-induced platelet aggregation. After removal of calcium, a small amount of procoagulant activity is present only in the void volume peak. These data suggest that both the procoagulant and von Willebrand activities are on the same molecule. Thus our previous conclusion remains the same: human factor VIII is a large glycoprotein composed of identical 195 000-molecular-weight subunits jointed by disulfide bonds and is responsible for both antihemophilic and von Willebrand activities in human plasma.